Team:ITESM-CEM/Interlab
From 2014.igem.org
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Relative Fluorescence Units (RFU) for each bacterial strain, as determined by triplicate with the fluorometer for each overnight culture and 2-hour subculture sample are reported at Appendix A. Further, a statistical mean, as well as a value of standard deviation, was calculated for each triplicate of measurements; this data is also summarized in Appendix A. | Relative Fluorescence Units (RFU) for each bacterial strain, as determined by triplicate with the fluorometer for each overnight culture and 2-hour subculture sample are reported at Appendix A. Further, a statistical mean, as well as a value of standard deviation, was calculated for each triplicate of measurements; this data is also summarized in Appendix A. | ||
- | The relative fluorescence of both negative controls (DH5α and Top 10 strains) with respect to the optical density is presented in dispersion plots (figure 2). A smooth negative association was found for both sets of samples. The negative slope of the curve indicates that the biomass may absorb some of the basal fluorescence.<br><br> | + | The relative fluorescence of both negative controls (DH5α and Top 10 strains) with respect to the optical density is presented in dispersion plots (figure 2). A smooth negative association was found for both sets of samples. The negative slope of the curve indicates that the biomass may absorb some of the basal fluorescence.<br><br></p> |
- | <img src="https://static.igem.org/mediawiki/2014/f/f9/Interlab_Results_%282%29-2.jpg" align="middle" hspace="10" BORDER=10><br> | + | <p class="centeredImage"><img src="https://static.igem.org/mediawiki/2014/f/f9/Interlab_Results_%282%29-2.jpg" align="middle" hspace="10" BORDER=10><br> |
<b>Figure 2.</b> Mean Relative Fluorescence plot for negative control samples taken from overnight cultures at increasing optical densities. All correlation coefficients are shown. | <b>Figure 2.</b> Mean Relative Fluorescence plot for negative control samples taken from overnight cultures at increasing optical densities. All correlation coefficients are shown. | ||
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The values of relative fluorescence for the DH5α negative control were then subtracted from the lectures obtained when analysing the modified bacterial strains. Dispersion plots for each genetic device built this way are presented in figure 3. An analysis of correlation reveals a strong positive association between relative fluorescence and OD600 in E. coli transformed with BioBrick BBa_I20260 (device 1) for both samples (overnight culture and 2-hour subculture). However, there appears to be no increase in fluorescence on the plots for BioBrick BBa_K823005 + BBa_E0240 (device 2) and BBa_K823012 + BBa_E0240 (device3). | The values of relative fluorescence for the DH5α negative control were then subtracted from the lectures obtained when analysing the modified bacterial strains. Dispersion plots for each genetic device built this way are presented in figure 3. An analysis of correlation reveals a strong positive association between relative fluorescence and OD600 in E. coli transformed with BioBrick BBa_I20260 (device 1) for both samples (overnight culture and 2-hour subculture). However, there appears to be no increase in fluorescence on the plots for BioBrick BBa_K823005 + BBa_E0240 (device 2) and BBa_K823012 + BBa_E0240 (device3). | ||
- | Finally, the curves of relative fluorescence for overnight culture and 2-hour subculture samples of bacteria expressing BioBrick BBa_I20260 are superposed in a single plot (figure 4). Correlation (R2) is kept almost constant; but there is a clear increase in the slope of the line when comparing the 37°C, 2-hour subculture with the regular overnight culture stored at 4°C. <br><br> | + | Finally, the curves of relative fluorescence for overnight culture and 2-hour subculture samples of bacteria expressing BioBrick BBa_I20260 are superposed in a single plot (figure 4). Correlation (R2) is kept almost constant; but there is a clear increase in the slope of the line when comparing the 37°C, 2-hour subculture with the regular overnight culture stored at 4°C. <br><br></p> |
- | <img src="https://static.igem.org/mediawiki/2014/9/90/Interlab_Results_%282%29-5.jpg" align="middle" hspace="10" BORDER=10><br> | + | <p class="centeredImage"><img src="https://static.igem.org/mediawiki/2014/9/90/Interlab_Results_%282%29-5.jpg" align="middle" hspace="10" BORDER=10><br> |
- | <b>Figure 3.</b> Mean Relative Fluorescence plot for E. coli DH5α transformed with each of the three genetic devices, samples taken from overnight cultures at increasing optical densities. The basal levels of fluorescence were subtracted. All correlation coefficients are shown.<br><br> | + | <b>Figure 3.</b> Mean Relative Fluorescence plot for E. coli DH5α transformed with each of the three genetic devices, samples taken from overnight cultures at increasing optical densities. The basal levels of fluorescence were subtracted. All correlation coefficients are shown.<br><br></p> |
- | <img src="https://static.igem.org/mediawiki/2014/5/56/Interlab_Results_%282%29-8.jpg" align="middle" hspace="10" BORDER=10><br> | + | <p class="centeredImage"><img src="https://static.igem.org/mediawiki/2014/5/56/Interlab_Results_%282%29-8.jpg" align="middle" hspace="10" BORDER=10><br> |
<b>Figure 4.</b> Comparative plot of Mean Relative Fluorescence for genetically modified E. coli DH5α strain, transformed with BioBrick BBa_I20260, at increasing optical densities. The upper curve belongs to a sample taken of the 2-hour subculture at 37°C in LB media; while the lower one was obtained directly from an overnight culture stored at 4°C. The basal levels of fluorescence emission were subtracted. All correlation coefficients are shown. | <b>Figure 4.</b> Comparative plot of Mean Relative Fluorescence for genetically modified E. coli DH5α strain, transformed with BioBrick BBa_I20260, at increasing optical densities. The upper curve belongs to a sample taken of the 2-hour subculture at 37°C in LB media; while the lower one was obtained directly from an overnight culture stored at 4°C. The basal levels of fluorescence emission were subtracted. All correlation coefficients are shown. | ||
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<a name="Annex"><u><h2>Annexes</h2></u></a> | <a name="Annex"><u><h2>Annexes</h2></u></a> | ||
<p style="text-align: justify; text-justify: inter-word;"> | <p style="text-align: justify; text-justify: inter-word;"> | ||
- | Measurements taken for 150 μl samples of overnight bacterial cultures after 2 hours of incubation at 4°C | + | Measurements taken for 150 μl samples of overnight bacterial cultures after 2 hours of incubation at 4°C</p> |
- | <img src="https://static.igem.org/mediawiki/2014/7/79/Table1_%282%29-8.jpg" width="600" height="550" align="middle" hspace="10" BORDER=10><br><br> | + | <p class="centeredImage"><img src="https://static.igem.org/mediawiki/2014/7/79/Table1_%282%29-8.jpg" width="600" height="550" align="middle" hspace="10" BORDER=10><br><br></p> |
- | Measurements taken for 150 μl samples of overnight cultures of negative controls, and E. coli DH5α transformed with BioBrick BBa_I20260. Samples were subcultured for 2 hours at 37°C and constant stirring.<br> | + | <p style="text-align: justify; text-justify: inter-word;">Measurements taken for 150 μl samples of overnight cultures of negative controls, and E. coli DH5α transformed with BioBrick BBa_I20260. Samples were subcultured for 2 hours at 37°C and constant stirring.<br></p> |
- | <img src="https://static.igem.org/mediawiki/2014/b/b7/Table2_%282%29-9.jpg" width="500" height="500" align="middle" hspace="10" BORDER=10><br><br> | + | <p class="centeredImage"><img src="https://static.igem.org/mediawiki/2014/b/b7/Table2_%282%29-9.jpg" width="500" height="500" align="middle" hspace="10" BORDER=10><br><br></p> |
- | Statistical means and standard deviations calculated for each triplicate of measurements of relative fluorescence, using samples taken from overnight cultures stored for 2 hours at 4°C. | + | <p style="text-align: justify; text-justify: inter-word;">Statistical means and standard deviations calculated for each triplicate of measurements of relative fluorescence, using samples taken from overnight cultures stored for 2 hours at 4°C.</p> |
- | <img src="https://static.igem.org/mediawiki/2014/9/9b/Table3_%282%29-10.jpg" width="700" height="250" align="middle" hspace="10" BORDER=10><br><br> | + | <p class="centeredImage"><img src="https://static.igem.org/mediawiki/2014/9/9b/Table3_%282%29-10.jpg" width="700" height="250" align="middle" hspace="10" BORDER=10><br><br></p> |
- | Statistical means and standard deviations calculated for each triplicate of measurements of relative fluorescence, using samples taken from E. coli DH5α transformed with BioBrick BBa_I20260, subcultured for 2 hours at 37°C and constant stirring.<br> | + | <p style="text-align: justify; text-justify: inter-word;">Statistical means and standard deviations calculated for each triplicate of measurements of relative fluorescence, using samples taken from E. coli DH5α transformed with BioBrick BBa_I20260, subcultured for 2 hours at 37°C and constant stirring.<br></p> |
- | <img src="https://static.igem.org/mediawiki/2014/2/29/Table4_%282%29-10.jpg" width="380" height="350" align="middle" hspace="10" BORDER=10> | + | <p class="centeredImage"><img src="https://static.igem.org/mediawiki/2014/2/29/Table4_%282%29-10.jpg" width="380" height="350" align="middle" hspace="10" BORDER=10> |
Revision as of 00:54, 14 October 2014
ITESM-CEM | Enzy7-K me |
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Interlab
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